CN101390870A - Ginsenoside Rb1 containing impurity ginsenoside Rc - Google Patents
Ginsenoside Rb1 containing impurity ginsenoside Rc Download PDFInfo
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Abstract
The invention discloses a standard ginsenoside Rb1 active ingredient, containing ginsenoside Rb1 with the content more than or equal to 90% and less than 100%, is also characterized in that the active ingredient contains ginsenoside Rc with the content more than 0% and less than or equal to 10%; or the content of impurity ginsenoside Rc is more than or equal to 0.05% and less than or equal to 6.5%; or the content of ginsenoside Rc is more than or equal to 0.1% and less than or equal to 5.0%; the content of impurity ginsenoside Rc is more than or equal to 0.1% and less than or equal to 3.0%; Pharmacological experiments show that the standard active ingredient of the application has good pharmacological effects and can be prepared into pharmaceutical preparations in Pharmacy.
Description
Technical field
The present invention relates to technical field of Chinese medicines, be specifically related to a kind of ginsenoside Rb of standard
1Effective ingredient.
The invention reside in the middle pharmaceutically active ingredient that a kind of standard is provided, promptly contain the ginsenoside Rb of the standard of impurity ginsenoside Rc
1
Background technology
The application of Chinese medicine has had history in several thousand, for the mankind healthy made great contribution, along with day by day going deep into that Chinese medicine uses, the safety that Chinese medicine is used has caused global concern, because the particularity of Chinese medicine is (such as complicated component, factors such as composition is more), the monitoring that causes carrying out adverse reaction of tcm has certain degree of difficulty, but healthy for extensive patients, the monitoring dynamics of untoward reaction is bound to strengthen greatly, though Chinese medicine has very big particularity, but as long as (new drug development) carries out deep research from the source, such as studying at the impurity of middle pharmaceutically active ingredient earlier, long-pending few journey is many, and it is numerous to conform to the principle of simplicity, the science of setting up that is bound to, the adverse reaction of tcm detection architecture of standard; In view of science and technology development now, research at Chinese medicine should be that the basis is studied with middle pharmaceutically active ingredient and its big impurity, the research of impurity in the middle pharmaceutically active ingredient is clear, particularly that content is bigger impurity is studied and quantitatively control, help us clearly to understand the material base of effective ingredient, after the medicine listing, when untoward reaction or side effect take place, help us and effectively follow the trail of root, effectively impurity or synergism impurity should quantitatively be controlled, invalid impurity should carry out content and limit, poisonous impurity or objectionable impurities should further be removed, perhaps content is limited to harmless scope (such as less than 10PPM), therefore, the impurity in the centering pharmaceutically active ingredient carries out deep research, has profound significance, help the modernization of Chinese medicine, help Chinese medicine to go to the world, help the healthy of the mankind more.
Ginsenoside Rb
1Be mainly derived from Araliaceae Panax Radix Ginseng, Radix Notoginseng, Radix Panacis Quinquefolii, Panax vietnamensis Ha et Grushv., Rhizoma Panacis Japonici, in the root of plants such as Cucurbitaceae Gynostemma Herb Gynostemmae Pentaphylli, edge fruit Herb Gynostemmae Pentaphylli, stem, leaf, the alabastrum; Ginsenoside Rb
1Have significantly and pharmacologically active widely, except that having the good pharmacologically active, further studies show that in recent years: ginsenoside Rb at treating cardiac and cerebral vascular diseases
1Energy activator RNA polymerase, RNA, the DNA protein of promotion brain cell and nerve centre cell can be repaired impaired brain cell; make brain cell regeneration, the cerebral cortex cell is had protective effect, can improve ability of thinking; improve memory, prompting ginsenoside Rb
1Has nootropics, anti senile dementia drug exploitation value; Under the effect of certain anaerobe of enteral, can produce CompandK, be called for short Rck, and Rck has tangible cancer suppressing action, prompting ginsenoside Rb
1Having the cancer therapy drug exploitation is worth; Mice ig ginsenoside Rb
1, by improving the androgen level, activate the No/cGMP path, and significantly improve mouse sexual function, prompting ginsenoside Rb
1Has exploitation sexual function improving drug value.
Ginsenoside Rb
1Many employing laboratory methods obtain a small amount of (the mg level is to the g level), such as methods such as preparative high-performance liquid chromatographic method, silica gel column chromatographies, are used for scientific research, but can't obtain (more than the kg level) ginsenoside Rb in batches
1Be used for patent medicine exploitation and suitability for industrialized production, so this is ginsenoside Rb
1So far the reason and the difficult point that are difficult to patent medicine.
Application number is 03127664.4 patent documentation " environment-friendly type ginsenoside Rb
1Height obtain volume production already divide from ", the document discloses and can obtain content greater than 95% ginsenoside Rb
1, adopting silica gel column chromatography method, this method involves great expense, and is difficult to suitability for industrialized production; Another subject matter of the document is not have the method for analyzing and testing to show ginsenoside Rb
1Content reach 95%, its credibility be worth to be suspected; Application number is patent documentation " the ginsenoside Rb of 03148803.x
1Preparation technology ", the method that the document discloses the method+silica gel column chromatography of employing Amberlyst process+silica gel column chromatography obtains ginsenoside Rb
1, this method is only applicable to laboratory research equally, is difficult to realize suitability for industrialized production; Another subject matter of the document is to obtain ginsenoside Rb
1Do not have purity, do not have the method for assay yet; The patent documentation of application number " 200610093610.6 " method of extraction separation ginsenoside monomer " a kind of from Folium Ginseng ", the document discloses the method for employing Amberlyst process+silica gel column chromatography and has separated the method that obtains different ginsenoside monomers, comprises ginsenoside Rb
1Its subject matter does not still have the requirement of purity, does not have content assaying method, and this process still adopted this complexity of column chromatography, cost height, can't suitability for industrialized production method.
In sum, obtaining ginsenoside Rb
1On the basis of effective ingredient, study, obtain standard active ingredient, have profound significance at its impurity impurity that particularly content is bigger.
Summary of the invention
For these reasons, our scientific research personnel carries out extensively and profoundly research by the ginsenoside that different genera, the different place of production, distinct methods are obtained, in that " content is more than or equal to 90% ginsenoside Rb
1, total impurities is smaller or equal to 10% " the basis on, to ginsenoside Rb
1Effective ingredient carried out deep research again, by performing creative labour, proved conclusively ginsenoside Rb
1In content bigger one of impurity be the Ginsenoside Rc, further research is quantitatively controlled the Ginsenoside Rc, obtains standard ginsenoside Rb
1Effective ingredient, can be directly as medicine material in market sale, the preparation raw material that also can be used as the medicament preparation uses, and like this, helps the standardization of raw material of Chinese medicine, helps the standardization that Chinese medicine preparation is produced; Simultaneously, our scientific research personnel is also at ginsenoside Rb
1Extraction and purification process study: adopting Amberlyst process, is raw material with the total saponins, separates obtaining ginsenoside Rb
1Crude product by recrystallization method, obtains ginsenoside Rb again
1, this process is simple, can realize suitability for industrialized production, the ginsenoside Rb that obtains
1Cost is not high, its content and impurity is analyzed ginsenoside Rb
1Content is more than or equal to 90%, in the impurity Ginsenoside Rc's content smaller or equal to 10%, conformance with standard ginsenoside Rb
1The requirement of effective ingredient; The application contains the ginsenoside Rb of impurity ginsenoside Rc
1Carry out the pharmacologically active experiment, find that it except that having effects such as good treatment cardiovascular and cerebrovascular disease, antitumor, also has good effect for reducing blood fat, with existing ginsenoside Rb
1(it is extremely low not contain Ginsenoside Rc or content) relatively has utmost point significant difference (P＜0.01) aspect effect for reducing blood fat.
The present invention is achieved through the following technical solutions.
The application is to provide a kind of ginsenoside Rb of standard
1, i.e. the middle pharmaceutically active ingredient of standard: effective ingredient is carried out content determine, impurity is carried out material is determined and content is determined; The application's standard ginsenoside Rb
1Effective ingredient can be used as the pharmaceutical preparation crude drug and uses, on the basis that meets the pharmaceutical preparation requirement, and the ginsenoside Rb of preparation
1Pharmaceutical preparation can be used for the treatment of human body diseases; The impurity that the application determines is the Ginsenoside Rc;
The ginsenoside Rb of the application's standard
1Effective ingredient is:
(1) a kind of ginsenoside Rb
1, ginsenoside Rb
1Content is characterized in that more than or equal to 90% and less than 100% impurity comprises the Ginsenoside Rc, and impurity ginsenoside Rc content is greater than 0 and smaller or equal to 10%.
(2) a kind of ginsenoside Rb
1, ginsenoside Rb
1Content is more than or equal to 90% and less than 100%, and wherein the content of impurity ginsenoside Rc is more than or equal to 0.05% and smaller or equal to 6.5%.
(3) a kind of ginsenoside Rb
1, ginsenoside Rb
1Content is more than or equal to 90% and less than 100%, and wherein the content of impurity ginsenoside Rc is more than or equal to 0.1% and smaller or equal to 5.0%.
(4) a kind of ginsenoside Rb
1, ginsenoside Rb
1Content is more than or equal to 90% and less than 100%, and wherein the content of impurity ginsenoside Rc is more than or equal to 0.1% and smaller or equal to 3.0%.
The Ginsenoside Rc is that content is one of bigger in the impurity in the above-mentioned standard active ingredient, by scientific experiments it is separated, and determines its structure by serial experiment again.
Above-mentioned standard active ingredient can be by Radix Ginseng, Radix Notoginseng, Radix Panacis Quinquefolii, Panax vietnamensis Ha et Grushv., Rhizoma Panacis Japonici, obtain total saponins in the root of plants such as Cucurbitaceae Gynostemma Herb Gynostemmae Pentaphylli, edge fruit Herb Gynostemmae Pentaphylli, stem, leaf, the alabastrum, carrying out purification through the high performance liquid chromatography preparation method again obtains, also can carry out monomer separation by silica gel column chromatography, high performance liquid chromatography control terminal point obtains, or the like;
The above-mentioned standard ginsenoside Rb of the application
1Effective ingredient also can be obtained by following method:
(1) gets Radix Notoginseng, Radix Ginseng, Radix Panacis Quinquefolii or Herb Gynostemmae Pentaphylli and extract the total saponins that obtains;
(2) total saponins is dissolved in water, and filters, and filtrate is passed through nonpolar or low pole macroporous adsorptive resins, washing, 40% ethanol elution, and eluent discards, and reuse 50% ethanol elution is collected 50% eluent, drying;
(3) dry thing filters with ethanol, 95% ethanol, n-butyl alcohol or dissolve with methanol, and filtrate adds or do not add organic solvent, and placement is separated out, and filtration, drying obtain ginsenoside Rb
1
Total saponins in the above-mentioned steps 1 can directly be bought by market or prepare according to scientific and technical literature;
The ginsenoside Rb that above-mentioned steps (3) obtains
1Method that can repeating step (3);
Described macroporous resin column is HPD-100, HPD-100A, HPD-300, HPD-400, HPD-400A, HPD-450, D101,1300-I, 1400 or AB-8.
Described organic solvent is one or both in acetone, the ethyl acetate.
Above-mentioned ginsenoside Rb
1Pharmaceutical preparation for the unit dose of active substance preparation.
Above-mentioned ginsenoside Rb
1Be the pharmaceutical preparation of active substance preparation, wherein unit dose is 1mg-5000mg.
Above-mentioned ginsenoside Rb
1Be the pharmaceutical preparation of active substance preparation, wherein unit dose is 5mg-2000mg.
Above-mentioned ginsenoside Rb
1Treat and/or prevent application in hyperlipidemia, cardiovascular and cerebrovascular disease, tumor, aging, the hypomnesis medicine in preparation.
One, detection method
Experimental technique:
Adopt high performance liquid chromatography to carry out check and analysis;
Chromatographic column: C
18Reversed phase chromatographic column;
Chromatographic condition and system suitability experiment: with the octadecylsilane chemically bonded silica is filler; Flow velocity 1.0ml/min; 25 ℃ of column temperatures; Detect wavelength 203nm; Number of theoretical plate is by ginsenoside Rb
1Meter should be not less than 2500; With water: acetonitrile volume ratio=33:67 is a mobile phase;
The preparation of reference substance solution: precision takes by weighing ginsenoside Rb
1Reference substance adds dissolve with methanol and shakes up in volumetric flask, and is diluted to scale;
The preparation of need testing solution: precision takes by weighing or measures sample and adds dissolve with methanol in the volumetric flask and shake up, and is diluted to scale;
Algoscopy: accurate respectively absorption reference substance solution and need testing solution respectively inject chromatograph of liquid, adopt by external standard method with calculated by peak area, promptly.
Annotate: the application's Radix Ginseng saponin Rb
1Reference substance is the statutory standards product, buys from Chinese pharmaceutical biological product evaluation and buys.
The application's standard ginsenoside Rb
1The check and analysis of effective ingredient
Get the application's standard ginsenoside Rb
1Effective ingredient, carry out the check and analysis experimental result according to above-mentioned experimental technique and see Table 1:
Table 1 ginsenoside Rb
1The sample determination result
Experiment conclusion: show the application's standard ginsenoside Rb by above-mentioned experiment
1In the effective ingredient, ginsenoside Rb
1Content is more than or equal to 90% and less than 100%, and the content of impurity ginsenoside Rc is greater than 0% and smaller or equal to 10%; Ginsenoside Rb
1Content is more than or equal to 90% and less than 100%, and the content of impurity ginsenoside Rc is more than or equal to 0.05% and smaller or equal to 6.5%; Ginsenoside Rb
1Content is more than or equal to 90% and less than 100%, and the content of impurity ginsenoside Rc is more than or equal to 0.1% and smaller or equal to 5.0%; Ginsenoside Rb
1Content is more than or equal to 90% and less than 100%, and the content of impurity ginsenoside Rc is more than or equal to 0.1% and smaller or equal to 3.0%.
Two, Ginsenoside Rc's structural identification data
Get the application's standard ginsenoside Rb
1Effective ingredient separates through silica gel column chromatography, in conjunction with high performance liquid chromatography liquid method, and the impurity that content is bigger, ginsenoside Rb
1Separate respectively, obtain the impurity monomer, carry out structural identification, the conclusive evidence data is as follows:
m.p：198℃～200℃。Mass spectrum ESI-MS:1101,1077,945,914,783,765,459.
1HNMR(400MHz，pyridine-d5)：0.72(3H，s，19-CH
3)，0.94(6H，s，30-CH
3，18-CH
3)，1.08(3H，s，29CH
3)，1.26(3H，s，21CH
3)1.61(6H，26-CH
3，27-CH
3)，1.61(6H，s，26CH
3，27CH
3)，1.66(3H，s，28CH
3)，4.90(1H，d，J＝7.7Hz，H-1’)，5.35(1H，d，J＝7.4Hz，H-1”)，5.11(1H，d，J＝7.7Hz，H-1”’)，4.96(1H，d，J＝1.4Hz，H-1””)。
13CNMR(400MHz，pyridine-d5)：39.5(C-1)，26.6(C-2)，89.0(C-3)，39.5(C-4)，56.4(C-5)，18.4(C-6)，35.1(C-7)，40.0(C-8)，50.2(C-9)，36.8(C-10)，30.7(C-11)，70.3(C-12)，49.4(C-13)，51.4(C-14)30.7(C-15)，26.6(C-16)，51.7(C-17)，16.4(C-18)，16.0(C-19)83.1(C-20)，22.2(C-21)，36.0(C-22)，23.1(C-23)，125.8(C-24)，130.8(C-25)，25.4(C-26)，17.6(C-27)，28.1(C-28)，16.4(C-29)，17.3(C-30)，104.8(C-1’)，83.0(C-2’)，77.4(C-3’)，71.6(C-4’)，77.6(C-5’)，62.8(C-6’)，105.4(C-1”)，76.4(C-2”)，78.8(C-3”)，71.6(C-4”)，78.0(C-5”)，62.6(C-6”)，97.9(C-1”)，74.8(C-2”’)，78.0(C-3”’)，71.6(C-4”’)，76.3(C-5”’)，63.0(C-6””)，109.8(C-1””)，83.3(C-2””)，78.7(C-3””)，85.9(C-4””)，62.7(C-5””)。Therefore Ginsenoside Rc's data basically identical in above data and the document [1] determines that chemical compound is the Ginsenoside Rc.Document 1: Su Jian etc., the saponin constituent research of Radix Panacis Quinquefolii, CHINA JOURNAL OF CHINESE MATERIA MEDICA, 2003,28 (9), 830 are produced in Jilin.
Experiment conclusion: show that by above-mentioned experiment one of big impurity of the application's content is the Ginsenoside Rc.
Three, pharmacological evaluation
Experiment 1
The research of blood lipid regulation effect
Experiment medicine: the application's standard ginsenoside Rb
1Effective ingredient (meeting of the qualification of the application's standard active ingredient) to impurity ginsenoside Rc; Commercially available ginsenoside Rb
1Monomer (Chinese pharmaceutical biological product is identified institute); XUESHUANTONG ZHUSHEYE.
Experimental technique: male SD rat, adaptability were fed after the week, were divided into normal control group, model group, experimental program group at random by body weight.Except that the distilled water of normal control group every mornings 8 point-9 a filling stomach 1.0ml/100g, all the other each groups are irritated stomach 1.0ml/100g lipomul, prescription is 30% Adeps Sus domestica, 10% cholesterol, 2% cholate, 0.2% propylthiouracil, 20% Tween 80,20% propylene glycol, 10% sucrose, with distilled water melt into 100ml, continuous 10 days.12 hours socket of the eye venous plexuses of fasting are got blood, detect serum TC, TG, HDL-C and LDL-C content, and divide into groups again by the serum TC level.After adjusting grouping, each is organized rat every morning and gives lipomul by above-mentioned modeling test method, emulsion formulations becomes: 20% Adeps Sus domestica, 5% cholesterol, 2% cholate, 0.2% propylthiouracil, 10% tween, 10% propylene glycol, 5% sucrose distilled water melt into 100ml, point-2 were pressed and were respectively organized dosage tail intravenously administrable afternoon 1, in 4 weeks of successive administration, dosage is 20mg/kg.After 4 weeks of administration finish, fasting 12 hours, femoral artery is got blood, and a part of blood is used for hemorheology and detects with 1% anticoagulant heparin (about 4ml).Another part blood separation serum is used for the detection of biochemical indicator.Duration of test claims body weight weekly one time, adjusts dosage by body weight, and measures food ration.4 weeks of administration are detected serum TC, TG, LDL-C and HDL-C level respectively, and experimental result sees Table 2
4 weeks of table 2 administration are to the influence of rat fat
Annotate: compare with matched group
*P＜0.01,
*P＜0.05;
Brief summary: show by above-mentioned experiment, contain the ginsenoside Rb of impurity ginsenoside Rc
1Effective ingredient is with the ginsenoside Rb that does not contain Ginsenoside Rc's impurity
1Effective ingredient relatively has the effect of better blood fat reducing, illustrates that impurity ginsenoside Rc is effective impurity, itself and ginsenoside Rb
1Effect and synergism with addition.
Experiment 2
Protective effect to rat cerebral infarction
Experiment medicine: the application's standard ginsenoside Rb
1Effective ingredient (meeting of the qualification of the application's standard active ingredient) to impurity ginsenoside Rc; Commercially available ginsenoside Rb
1Monomer (Chinese pharmaceutical biological product is identified institute); XUESHUANTONG ZHUSHEYE.
Laboratory animal: SD rat, 250-300g, male and female half and half.
Experimental technique: get rat, chloral hydrate 300mg/kgip anesthesia, cervical incision, separate and the ligation right carotid, behind the suture muscles skin, right lateral position is fixed, and cuts skin at auris dextra and right eye outer canthus line mid point, separate temporalis, expose zygomatic process and temporal bone, open the bone window of about a 3 * 3mm at head end 1～2mm place of zygomatic process, expose middle cerebral artery (MCA), it is disconnected that MCA is burnt, and sews up the incision.With reference to the administration of Bederson method, put to death animal, get right cerebral hemisphere, be cut into 5, place 37 ℃ of incubation 15min dyeing of 2g/LNPT, the white infarct cerebral tissue that carefully takes and weigh and do not dyed blue color, experimental result sees Table 3.
Table 3 different pharmaceutical is protected situation to rat cerebral infarction
Annotate: compare with the normal saline group
*P＜0.01; Compare #P＜0.05 with the XUESHUANTONG ZHUSHEYE group
Experiment 3
To the protective effect of Cor Canitis myocardial ischemia
Laboratory animal: Beagle Canis familiaris L., body weight 7.5-10kg.
Experiment medicine: the application's standard ginsenoside Rb
1Effective ingredient; Commercially available ginsenoside Rb
1Monomer (Chinese pharmaceutical biological product is identified institute); XUESAITONG ZHUSHEYE.
Experimental technique: get experimental dogs, be divided into normal saline group, XUESAITONG ZHUSHEYE group, the application's standard ginsenoside Rb
1Effective ingredient group, commercially available ginsenoside Rb
1Set of monomers, the administration group is respectively in the subcutaneous cephalic vein administration of Canis familiaris L. forelimb, the each 20mg/kg of dosage, normal saline groups etc. are held inequality, administration every day 1 time, successive administration 3 days, administration are used pentobarbital sodium 25mg/kg intravenous anesthesia, tracheal intubation after the 3rd day, connect artificial respirator positive pressure respiration in addition, the 5th rib is opened breast from the left side, cuts off pericardium, exposes heart, the pericardial incision edge is sewn in thoracic wall, divide the second stage of ligation at ramus descendens anterior arteriae coronariae sinistrae, and slow iv lignocaine 8mg/kg is in case the ventricular fibrillation that may cause before the ligation is sewed up pericardium and thoracic wall subsequently.24h behind the heart infarction, Canis familiaris L. is put to death after pentobarbital sodium anesthesia, take out heart rapidly, excision trunk and fatty tissue, check the position of anterior descending branch ligation point, claim heavy whole-heartedly, excise right ventricle and left atrium then, stay interventricular septum and left atrium, claim to such an extent that left ventricle is heavy, from the apex of the heart and beginning and anterior descending branch vertical direction left ventricle is cut into the thick flesh sheet of 2-3mm, be dipped in dyeing 30min in the nitro tetrazolium blue solution (NBT), cut ischemic region cardiac muscular tissue and weigh, the calculating myocardium infarction size, that is: heart infarction scope=ischemic region weight/left ventricle is heavy by * 100%, the results are shown in Table 4.
The influence of table 4 pair Cor Canitis chamber infarction size
Annotate: compare with the normal saline group,
*P＜0.01; Compare #P＜0.05 with positive controls XUESAITONG ZHUSHEYE group
Experiment 4
Comparison to the rats'liver tumor suppression
Laboratory animal: rat, 150g-180g, male and female are regardless of.
Experiment medicine: normal saline; The application's standard ginsenoside Rb
1Effective ingredient; Commercially available ginsenoside Rb
1Monomer (Chinese pharmaceutical biological product is identified institute); Ad pro injection (Guizhou Yibai Pharmaceutical Co., Ltd)
Experimental technique: get rat and be divided into normal saline group, commercially available ad pro injection group, the application's standard ginsenoside Rb
1Effective ingredient group, commercially available ginsenoside Rb
1Set of monomers is made W
256Liver in inoculation, inoculate after 7 days, press the dosage intraperitoneal injection of anesthesia of 35mg/kg with pentobarbital sodium, fixing, cutting open the belly exposes liver, tumor surface maximum diameter (a) and path (b) are pressed (a*b on the measurement liver
2)/2=V (gross tumor volume).Separate stomach, arteria duodenalis, common hepatic artery and proper hepatic artery, ligation stomach, arteria duodenalis far-end, with silver brain clip blocking-up common hepatic artery, in sending into proper hepatic artery again at the gastroduodenal artery upper cut and after inserting external diameter 0.3mm conduit under the operating loupe, inject respectively by the experiment grouping then and be subjected to reagent thing (dosage is identical), postoperative tube drawing ligation gastroduodenal artery, decontrol the common hepatic artery silver brain clip, sew up the incision again, place animal housing to wait to revive rat, continue breeding observing, performed the operation back 8 days, detect gross tumor volume by last method, experimental result sees Table 5:
Table 5 is respectively organized preparation to the rejection ratio of tumor
Annotate: compare with the normal saline group
*P＜0.01; Compare #P＜0.05 with the commercially available ad pro injection group of positive controls
Experiment 5
Physics, chemistry and biological factor all can be brought out cancer, but the 80%-90% of people's cancer pathogenic factor thinks what the Environmental Chemistry material caused.
Diethylnitrosamine (DENA) is brought out the inhibition of rat liver cancer
Laboratory animal: rat, about 120g, male and female are regardless of.
Experiment medicine: normal saline; The application's standard ginsenoside Rb
1Effective ingredient; Commercially available ginsenoside Rb
1Monomer (Chinese pharmaceutical biological product is identified institute); Ad pro injection (Guizhou Yibai Pharmaceutical Co., Ltd) experimental technique: feed rat with the drinking-water that contains diethylnitrosamine (DENA), 36 all primary hepatocarcinoma that get experimentize according to the method for testing 3, record survival of rats natural law, and experimental result sees Table 6:
Table 6 survival of rats natural law relatively
Annotate: compare with the normal saline group
*P＜0.01; Compare #P＜0.05 with the commercially available ad pro injection group of positive controls
Annotate: with the application's standard ginsenoside Rb
1Effective ingredient and commercially available ginsenoside Rb
1Monomer carries out experiment, dementia experiment, the sexual function improving experiment of mouse memory function, and experimental result shows, the application's standard ginsenoside Rb
1Effective ingredient is than commercially available ginsenoside Rb
1The monomer pharmacologically active is good.
Discuss: show that by above-mentioned pharmacological evaluation the application contains the ginsenoside Rb of impurity ginsenoside Rc
1Ratio content is near 100% ginsenoside Rb
1The monomer pharmacologically active will be got well, and through our research, primary explanation is: the Ginsenoside Rc is an active component, and the two except that having summation action, also can have synergism in entering body, in addition, contain the ginsenoside Rb of a certain amount of impurity ginsenoside Rc
1Aspect drug metabolism, can promote ginsenoside Rb
1Absorption, and content is near 100% ginsenoside Rb
1May be not contain Ginsenoside Rc or content trace, to ginsenoside Rb
1Influence very little, can not produce synergism (aspect such as pharmacology and medicine generation), therefore, its pharmacologically active is than the application's standard ginsenoside Rb
1The pharmacologically active of effective ingredient is low.
Four, dose screening experiment
Protective effect to the anesthetized rat myocardial ischemia reperfusion injury
Experimental technique:
Get the healthy SD rat, body weight 240-260g, random packet: blank group, sham operated rats, model group, diltiazem hydrochloride injection matched group, ginsenoside Rb
1The various dose group.Place the pre-raising of equivalent environment 2 days, free diet.After pre-raising finishes, experimentize, animal is weighed, and 20% urethane is pressed the 0.6ml/100g lumbar injection, after treating that anesthesia is satisfied, lie on the back and be fixed on the Mus plate, tracheal intubation connects respirator, by 10～12ml tidal volume, 70 times/minute frequency is exhaled, and continuous positive pressure breathing is inhaled: exhale than being 1: 1.Adjust respiration parameter according to the respiratory frequency and the degree of depth.Connect electrocardiograph subsequently, survey normal ECG.Cut off front field of operation hair, iodine disinfection, cut off skin, subcutaneous tissue, front muscle and fascia 3～4cm, it is long to separate Intercostal muscle 3cm with the 18# vascular forceps along the 3rd intercostal passivity, open thoracic cavity and pericardium, recording ecg, strut 3,4 ribs, refer to hold thoracic cavity, rat right side with left hand four, the assistant upwards pushes away thymus with the ophthalmology tweezer, finds ligation sign blood vessel great cardiac vein between left auricle and pulmonary conus, 2mm place noinvasive roundlet pin band 6-0 silk thread threading below left auricle, depth of needle is 1～1.5mm, wide 2～3mm, recording ecg behind the threading, give corresponding medicinal liquid through the tail vein, recording ecg behind the administration 10min, and with one the band groove little plastics pipe pad at the ligation position, the ligation thereon of two rear line heads.At once recording ecg after the ligation is cyanosis or the II S-T section back of a bow that leads with left chamber antetheca and upwards raises greater than 0.1mv and be that ligation successfully indicates (it is superseded that the S-T section does not have the changer) more than the lasting 0.5h.10min recording ecg is once more cut off ligature behind the ligation 30min after the ligation, realizes perfusion again, and record pours into electrocardiogram at once again, removes in the thoracic cavity layer-by-layer suture thoracic wall behind the hematocele, removes respirator, animal recovery autonomous respiration, and incision of trachea does not process.Irritate again at once, 10min, 20min, 40min, 1h, 2h, 3h recording ecg respectively.Irritated again 3 hours, through abdominal aortic blood, 4000rpm is centrifugal, and 10min gets serum, adopt automatic clinical chemistry analyzer to detect LDH, CK and CK-MB activity, and adopt the corresponding reagent box to detect SOD in serum, MDA, dissection is cored dirty, the residual blood of ice normal saline flush away, cut off atrium and right ventricle, put into refrigerator and cooled immediately and freeze.With heart after refrigerator and cooled is frozen 10min, from the apex of the heart entad the parallel coronary sulcus direction in the end 5 of equal thickness are cut in left chamber, put into the 1%TTC dye liquor, 37 ℃ of dyeing 10min, the necrotic area is not a kermesinus, the necrotic area is canescence.Digital camera is taken pictures.Weighed respectively in necrotic area and non-necrotic area, calculate the percentage ratio that the necrotic area accounts for left ventricular mass, i.e. infarction size.
The detection index is respectively, myocardial infarct size.Experimental result sees table 6 for details:
The influence of myocardial ischemia myocardial infarct size (%) due to table 6 pair ligation/logical again rat ramus descendens anterior arteriae coronariae sinistrae (x ± s)
Annotate: compare with model group,
*P＜0.05,
*P＜0.01,
* *P＜0.001.
Experiment conclusion: show by the dose screening experiment, the application's Radix Ginseng saponin Rb1 increases pharmacologically active with dosage and also strengthens, it is different with the pharmacologically active of a lot of medicines that (pharmacologically active can arrive a plateau to most of medicine with the dosage increase, be that dosage increases again and activity does not increase), along with increasing activity, dosage increases always, do not have plateau, can arrive always and occur till the toxicity; Through us dose screening experiment and toxicity test, determine that the unit dose of the application's Radix Ginseng saponin Rb1 is 1mg-5000mg; Preferred unit dosage is 5mg-2000mg (dosage toxic reaction occurs greater than 5000mg, does not have significant difference less than 1mg and model group).
The application's standard ginsenoside Rb
1Effective ingredient and prior art ginsenoside Rb
1(" application number is patent documentation＜environment-friendly type ginsenoside Rb of 03127664.4
1Height obtain volume production already divide from, the document discloses and can obtain content greater than 95% ginsenoside Rb
1" etc. method obtain) relatively have following characteristics:
1, the application is with ginsenoside Rb
1In one of bigger impurity of content ownership of carrying out, determined that it is the Ginsenoside Rc, further clear and definite material base, and just obtained the ginsenoside Rb of certain content in the prior art
1, not carrying out deep research, material base is fuzzy;
2, the application marks ginsenoside Rb
1Effective ingredient can be used as the raw material of pharmaceutical preparation, in clinical practice, any untoward reaction or side effect take place, because material base is clearer and more definite, particularly impurity clearly can be in the research of follow-up clinical drug, go out clearly that the composition of medicine own causes, or the no related substance of introducing in the preparation of pharmaceutical formulations process (such as factors such as thermal source, pollutions) causes, help the development of medicine, and ginsenoside Rb of the prior art
1Though be effective ingredient, also exist certain camera bellows, when untoward reaction or side effect particularly take place, can't demand origin, cause the obstacle (such as the Herba Houttuyniae injectio incident) of drug development;
3, the application determines that impurity ginsenoside Rc belongs to effective impurity under study for action, on certain basis to effective ingredient ginsenoside Rb
1Have synergism, belong to effective impurity, and the prior art ginsenoside Rb that has been clear and definite
1Content, all the other materials are belonged to effective impurity, invalid impurity or objectionable impurities know nothing, research belongs to initial period.
Four, embodiment
Embodiment 1
A kind of ginsenoside Rb
1, ginsenoside Rb
1Content 90.02%, impurity content 9.98%, impurity comprises the Ginsenoside Rc, Ginsenoside Rc's content is 9.95.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 2
A kind of ginsenoside Rb
1, ginsenoside Rb
1Content 99.80%, impurity content 0.20%, impurity comprises the Ginsenoside Rc, Ginsenoside Rc's content is 0.13%.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 3
A kind of ginsenoside Rb
1, ginsenoside Rb
1Content 91.82%, impurity content 8.18%, impurity comprises the Ginsenoside Rc, Ginsenoside Rc's content is 8.09%.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 4
A kind of ginsenoside Rb
1, ginsenoside Rb
1Content 92.27%, the content 6.50% of impurity ginsenoside Rc.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 5
A kind of ginsenoside Rb
1, ginsenoside Rb
1Content 97.41%, the content 0.05% of impurity ginsenoside Rc.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 6
A kind of ginsenoside Rb
1, ginsenoside Rb
1Content 99.19%, the content 0.08% of impurity ginsenoside Rc.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 7
A kind of ginsenoside Rb
1, ginsenoside Rb
1Content 93.24%, the content 5.68% of impurity ginsenoside Rc.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 8
A kind of ginsenoside Rb
1, ginsenoside Rb
1Content 94.89%, the content 3.62% of impurity ginsenoside Rc.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 9
A kind of ginsenoside Rb
1, ginsenoside Rb
1Content 94.11%, the content 5.00% of impurity ginsenoside Rc.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 10
A kind of ginsenoside Rb
1, ginsenoside Rb
1Content 99.47%, the content 0.10% of impurity ginsenoside Rc.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 11
A kind of ginsenoside Rb
1, ginsenoside Rb
1Content 95.34%, the content 4.41% of impurity ginsenoside Rc.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 12
A kind of ginsenoside Rb
1, ginsenoside Rb
1Content 97.02%, the content 2.58% of impurity ginsenoside Rc.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 13
A kind of ginsenoside Rb
1, ginsenoside Rb
1Content 95.20%, the content 3.00% of impurity ginsenoside Rc.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 14
A kind of ginsenoside Rb
1, ginsenoside Rb
1Content 97.81%, the content 1.58% of impurity ginsenoside Rc.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
The Ginsenoside Rc is that content is one of bigger in the impurity in the foregoing description 1-14 standard active ingredient, by scientific experiments it is separated, and determines its structure by serial experiment again.
The foregoing description 1-14 standard active ingredient can be by Radix Ginseng, Radix Notoginseng, Radix Panacis Quinquefolii, Panax vietnamensis Ha et Grushv., Rhizoma Panacis Japonici, obtain total saponins in the root of plants such as Cucurbitaceae Gynostemma Herb Gynostemmae Pentaphylli, edge fruit Herb Gynostemmae Pentaphylli, stem, leaf, the alabastrum, carrying out purification through the high performance liquid chromatography preparation method again obtains, also can carry out monomer separation by silica gel column chromatography, high performance liquid chromatography control terminal point obtains, or the like; Also can prepare by following embodiment method.
Embodiment 15
Get Radix Ginseng and extract the Radix Ginseng total saponins obtain and be dissolved in water, filter, filtrate is by the HPD-100A macroporous adsorptive resins, washing, 40% ethanol elution, eluent discards, reuse 50% ethanol elution, collect 50% eluent, drying, dry thing 95% dissolve with ethanol filters, filtrate adds acetone, placement is separated out, and filtration, drying obtain ginsenoside Rb
1
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 16
Get Radix Notoginseng and extract the Radix Notoginseng total arasaponins obtain and be dissolved in water, filter, filtrate is by the HPD-100 macroporous adsorptive resins, washing, 40% ethanol elution, eluent discards, reuse 50% ethanol elution, collect 50% eluent, drying, dry thing 95% dissolve with ethanol filters, filtrate adds the organic solvent ethyl acetate, placement is separated out, and filtration, drying obtain ginsenoside Rb
1
Described Radix Ginseng total saponins extracting method prepares according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 17
Get Herb Gynostemmae Pentaphylli and extract and to obtain total saponins and be dissolved in water, filter, filtrate is by 1400 macroporous adsorptive resins, washing, 40% ethanol elution, eluent discards, reuse 50% ethanol elution, collect 50% eluent, drying, dry thing dissolves with n-butyl alcohol, filters, filtrate adds organic solvent-acetone, ethyl acetate, placement is separated out, and filtration, drying obtain ginsenoside Rb
1
Described Herb Gynostemmae Pentaphylli total glycosides extracting method prepares according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 18
Get Radix Panacis Quinquefolii and extract the total saponins obtain and be dissolved in water, filter, filtrate is by 1300-I macroporous adsorptive resins, washing, 40% ethanol elution, eluent discards, reuse 50% ethanol elution, collect 50% eluent, drying, dry thing dissolve with methanol filters, filtrate adds organic solvent-acetone, placement is separated out, and filtration, drying obtain ginsenoside Rb
1
The extracting method of described American ginseng total saponins can lead to the existing patent documentation of basis or scientific and technical literature prepares.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 19
Get Herb Gynostemmae Pentaphylli total glycosides and be dissolved in water, filter, filtrate is by 1400 macroporous adsorptive resins, washing, 40% ethanol elution, eluent discards, reuse 50% ethanol elution, collect 50% eluent, drying, dry thing dissolve with ethanol filters, filtrate adds the organic solvent ethyl acetate, placement is separated out, and filtration, drying obtain ginsenoside Rb
1
Described extracting gypenosides can be led to the existing patent documentation of basis or scientific and technical literature prepares.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 20
Get Radix Ginseng total saponins and be dissolved in water, filter, filtrate is by the D101 macroporous adsorptive resins, washing, 40% ethanol elution, eluent discards, reuse 50% ethanol elution, collect 50% eluent, drying, dry thing 95% dissolve with ethanol filters, filtrate adds organic solvent-acetone, placement is separated out, and filtration, drying obtain ginsenoside Rb
1
The extracting method of described Radix Ginseng total saponins can lead to the existing patent documentation of basis or scientific and technical literature prepares.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 21
Get Radix Notoginseng total arasaponins and be dissolved in water, filter, filtrate is by HPD-450 macroporous adsorptive resins, washing, 40% ethanol elution, eluent discards, reuse 50% ethanol elution, collect 50% eluent, drying, dry thing dissolves with n-butyl alcohol, filters, filtrate adds organic solvent-acetone, ethyl acetate, placement is separated out, and filtration, drying obtain ginsenoside Rb
1
Described Radix Notoginseng total arasaponins is by commercially available purchase.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 22
Get Radix Notoginseng total arasaponins and be dissolved in water, filter, filtrate is by the AB-8 macroporous adsorptive resins, washing, 40% ethanol elution, eluent discards, reuse 50% ethanol elution, collect 50% eluent, drying, dry thing dissolve with methanol filters, filtrate adds the organic solvent ethyl acetate, placement is separated out, and filtration, drying obtain ginsenoside Rb
1
Described Radix Notoginseng total arasaponins according to existing patent documentation or scientific and technical literature prepare (Ou Lailiang, Shi Zuoqing, Shi Rongfu, etc., the epistasis macroporous adsorbent resin is to the separation and purification research of arasaponin, Chinese herbal medicine, 2003,34 (10): 905-907; ).
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 23
Get Radix Panacis Quinquefolii and extract the total saponins obtain and be dissolved in water, filter, filtrate is by the HPD-300 macroporous adsorptive resins, washing, 40% ethanol elution, eluent discards, reuse 50% ethanol elution, collect 50% eluent, drying, dry thing 95% dissolve with ethanol filters, filtrate adds the organic solvent ethyl acetate, placement is separated out, and filtration, drying obtain ginsenoside Rb
1
Described American ginseng total saponins extracting method prepares according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 24
Get Radix Notoginseng total arasaponins and be dissolved in water, filter, filtrate is by the AB-8 macroporous adsorptive resins, washing, 40% ethanol elution, eluent discards, reuse 50% ethanol elution, collect 50% eluent, drying, dry thing 95% dissolve with ethanol filters, filtrate adds organic solvent-acetone, placement is separated out, and filtration, drying obtain ginsenoside Rb
1
Described Radix Notoginseng total arasaponins according to existing patent documentation or scientific and technical literature prepare (Yang Chongren, kingdom swallow, 5 jewels, etc., the saponin component of Radix Notoginseng reed head, pharmacy circular, 1985,20 (6): 337-338).
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 25
Get Radix Ginseng and extract and to obtain total saponins and be dissolved in water, filter, filtrate is by the HPD-400A macroporous adsorptive resins, washing, 40% ethanol elution, eluent discards, reuse 50% ethanol elution, collect 50% eluent, drying, dry thing dissolves with n-butyl alcohol, filters, filtrate adds the organic solvent ethyl acetate, placement is separated out, and filtration, drying obtain ginsenoside Rb
1
Described Radix Ginseng total saponins extracting method prepares according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 26
Get Radix Notoginseng and extract and to obtain total saponins and be dissolved in water, filter, filtrate is by the HPD-400 macroporous adsorptive resins, washing, 40% ethanol elution, eluent discards, reuse 50% ethanol elution, collect 50% eluent, drying, dry thing dissolve with methanol filters, filtrate adds the organic solvent ethyl acetate, placement is separated out, and filtration, drying obtain ginsenoside Rb
1
Described Radix Notoginseng total arasaponins prepares according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 27
Get Radix Notoginseng and extract the total saponins obtain and be dissolved in water, filter, filtrate is by the HPD-100 macroporous adsorptive resins, washing, 40% ethanol elution, eluent discards, reuse 50% ethanol elution, collect 50% eluent, drying, dry thing dissolve with methanol filters, filtrate adds organic solvent-acetone, placement is separated out, and filtration, drying obtain ginsenoside Rb
1
Described Radix Notoginseng total arasaponins according to existing patent documentation or scientific and technical literature prepare (Yang Chongren, kingdom swallow, 5 jewels, etc., the saponin component of Radix Notoginseng reed head, pharmacy circular, 1985,20 (6): 337-338; ).
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 28
Get Radix Ginseng and extract and to obtain Radix Ginseng total saponins and be dissolved in water, filter, filtrate is by the HPD-450 macroporous adsorptive resins, washing, 40% ethanol elution, eluent discards, reuse 50% ethanol elution, collect 50% eluent, drying, dry thing dissolve with ethanol filters, filtrate adds the organic solvent ethyl acetate, placement is separated out, and filtration, drying obtain ginsenoside Rb
1
Described Radix Ginseng total saponins extracting method prepares according to existing literature method;
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 29
Get Radix Panacis Quinquefolii and extract the American ginseng total saponins obtain and be dissolved in water, filter, filtrate is by the HPD-100A macroporous adsorptive resins, washing, 40% ethanol elution, eluent discards, and reuse 50% ethanol elution is collected 50% eluent, dry, dry thing dissolve with ethanol filters, and filtrate placement is separated out, filtration, drying obtain ginsenoside Rb
1
Described American ginseng total saponins is by commercially available purchase;
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 30
Get Radix Ginseng and extract the Radix Ginseng total saponins obtain and be dissolved in water, filter, filtrate is by the HPD-100 macroporous adsorptive resins, washing, 40% ethanol elution, eluent discards, and reuse 50% ethanol elution is collected 50% eluent, dry, dry thing 95% dissolve with ethanol filters, and filtrate placement is separated out, filtration, drying obtain ginsenoside Rb
1
Described Radix Ginseng total saponins extracting method prepares according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 31
Get Radix Notoginseng and extract and to obtain total saponins and be dissolved in water, filter, filtrate is by the D101 macroporous adsorptive resins, washing, 40% ethanol elution, eluent discards, and reuse 50% ethanol elution is collected 50% eluent, dry, dry thing dissolves with n-butyl alcohol, filters, and filtrate placement is separated out, filtration, drying obtain ginsenoside Rb
1
Described Radix Notoginseng total arasaponins extracting method prepares according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 32
Get Radix Panacis Quinquefolii and extract the total saponins obtain and be dissolved in water, filter, filtrate is by 1300-I macroporous adsorptive resins, washing, 40% ethanol elution, eluent discards, and reuse 50% ethanol elution is collected 50% eluent, dry, dry thing dissolve with methanol filters, and filtrate placement is separated out, filtration, drying obtain ginsenoside Rb
1
The extracting method of described American ginseng total saponins can lead to the existing patent documentation of basis or scientific and technical literature prepares.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 33
Get Radix Ginseng total saponins and be dissolved in water, filter, filtrate is by 1400 macroporous adsorptive resins, washing, 40% ethanol elution, eluent discards, and reuse 50% ethanol elution is collected 50% eluent, dry, dry thing dissolve with ethanol filters, and filtrate placement is separated out, filtration, drying obtain ginsenoside Rb
1
Described Radix Ginseng total saponins prepares according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 34
Get Radix Notoginseng total arasaponins and be dissolved in water, filter, filtrate is by the AB-8 macroporous adsorptive resins, washing, 40% ethanol elution, eluent discards, and reuse 50% ethanol elution is collected 50% eluent, dry, dry thing 95% dissolve with ethanol filters, and filtrate placement is separated out, filtration, drying obtain ginsenoside Rb
1
Described Radix Notoginseng total arasaponins prepares according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 35
Get Herb Gynostemmae Pentaphylli and extract and to obtain total saponins and be dissolved in water, filter, filtrate is by the HPD-450 macroporous adsorptive resins, washing, 40% ethanol elution, eluent discards, and reuse 50% ethanol elution is collected 50% eluent, dry, dry thing dissolves with n-butyl alcohol, filters, and filtrate placement is separated out, filtration, drying obtain ginsenoside Rb
1
Described Herb Gynostemmae Pentaphylli total glycosides extracting method prepares according to existing document.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 36
Get Radix Ginseng and extract and to obtain total saponins and be dissolved in water, filter, filtrate is by the HPD-400 macroporous adsorptive resins, washing, 40% ethanol elution, eluent discards, and reuse 50% ethanol elution is collected 50% eluent, dry, dry thing dissolve with methanol filters, and filtrate placement is separated out, filtration, drying obtain ginsenoside Rb
1
Described Radix Ginseng total saponins prepares according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 37
Get Radix Notoginseng and extract the total saponins obtain and be dissolved in water, filter, filtrate is by the HPD-400A macroporous adsorptive resins, washing, 40% ethanol elution, eluent discards, and reuse 50% ethanol elution is collected 50% eluent, dry, dry thing 95% dissolve with ethanol filters, and filtrate placement is separated out, filtration, drying obtain ginsenoside Rb
1
Described Radix Notoginseng total arasaponins extracting method prepares according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 38
Get American ginseng total saponins and be dissolved in water, filter, filtrate is by the HPD-100 macroporous adsorptive resins, washing, 40% ethanol elution, eluent discards, and reuse 50% ethanol elution is collected 50% eluent, dry, dry thing 95% dissolve with ethanol filters, and filtrate placement is separated out, filtration, drying obtain ginsenoside Rb
1
Described American ginseng total saponins prepares according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 39
Get Herb Gynostemmae Pentaphylli and extract and to obtain total saponins and be dissolved in water, filter, filtrate is by the HPD-300 macroporous adsorptive resins, washing, 40% ethanol elution, eluent discards, and reuse 50% ethanol elution is collected 50% eluent, dry, dry thing dissolves with n-butyl alcohol, filters, and filtrate placement is separated out, filtration, drying obtain ginsenoside Rb
1
Described Herb Gynostemmae Pentaphylli total glycosides extracting method prepares according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 40
Get Radix Ginseng and extract and to obtain total saponins and be dissolved in water, filter, filtrate is by the AB-8 macroporous adsorptive resins, washing, 40% ethanol elution, eluent discards, and reuse 50% ethanol elution is collected 50% eluent, dry, dry thing dissolve with methanol filters, and filtrate placement is separated out, filtration, drying obtain ginsenoside Rb
1
Described Radix Ginseng total saponins prepares according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 41
Get Radix Notoginseng total arasaponins and be dissolved in water, filter, filtrate is by 1400 macroporous adsorptive resins, washing, 40% ethanol elution, eluent discards, and reuse 50% ethanol elution is collected 50% eluent, dry, dry thing dissolve with methanol filters, and filtrate placement is separated out, filtration, drying obtain ginsenoside Rb
1
Described Radix Notoginseng total arasaponins prepares according to existing patent documentation or scientific and technical literature.
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
Embodiment 42
Get Radix Ginseng total saponins and be dissolved in water, filter, filtrate is by the HPD-450 macroporous adsorptive resins, washing, 40% ethanol elution, eluent discards, and reuse 50% ethanol elution is collected 50% eluent, dry, dry thing dissolve with ethanol filters, and filtrate placement is separated out, filtration, drying obtain ginsenoside Rb
1
Described Radix Ginseng total saponins is by commercially available purchase;
Above-mentioned ginsenoside is as Rb
1Medicine material can be prepared into acceptable pharmaceutical preparation on the pharmaceutics.
The foregoing description 15-42 obtains ginsenoside Rb
1In the effective ingredient, ginsenoside Rb
1Content is more than or equal to 90% and less than 100%, and the content of impurity ginsenoside Rc is greater than 0% and smaller or equal to 10%; Perhaps ginsenoside Rb
1Content is more than or equal to 90% and less than 100%, and the content of impurity ginsenoside Rc is more than or equal to 0.05% and smaller or equal to 6.5%; Perhaps ginsenoside Rb
1Content is more than or equal to 90% and less than 100%, and the content of impurity ginsenoside Rc is more than or equal to 0.1% and smaller or equal to 5.0%; Perhaps ginsenoside Rb
1Content is more than or equal to 90% and less than 100%, and the content of impurity ginsenoside Rc is more than or equal to 0.1% and smaller or equal to 3.0%.
The ginsenoside Rb of the foregoing description 1-44
1Be the pharmaceutical preparation of active substance preparation, wherein unit dose is 1mg-5000mg.
The foregoing description 1-44 above-mentioned ginsenoside Rb
1Be the pharmaceutical preparation of active substance preparation, wherein unit dose is 5mg-2000mg.
Above-mentioned ginsenoside Rb
1Treat and/or prevent application in cardiovascular and cerebrovascular disease, tumor, aging, the hypomnesis medicine in preparation.
Annotate: the present invention's concrete technical scheme required for protection is not limited to the concrete combination of the expressed technical scheme of the foregoing description.
Claims (12)
1, a kind of ginsenoside Rb
1, ginsenoside Rb
1Content is more than or equal to 90% and less than 100%, it is characterized in that also containing content greater than 0% and smaller or equal to 10% Ginsenoside Rc.
2, a kind of ginsenoside Rb according to claim 1
1, wherein Ginsenoside Rc's content is more than or equal to 0.05% and smaller or equal to 6.5%.
3, a kind of ginsenoside Rb according to claim 1
1, wherein Ginsenoside Rc's content is more than or equal to 0.1% and smaller or equal to 5.0%.
4, a kind of ginsenoside Rb according to claim 1
1, wherein Ginsenoside Rc's content is more than or equal to 0.1% and smaller or equal to 3.0%.
5, according to each described a kind of ginsenoside Rb of claim 1-4
1, its preparation method is: get Radix Notoginseng, Radix Ginseng, Radix Panacis Quinquefolii or Herb Gynostemmae Pentaphylli and extract the total saponins that obtains, total saponins is dissolved in water, filter, filtrate is passed through nonpolar or low pole macroporous adsorptive resins, washing, 40% ethanol elution, eluent discards, and reuse 50% ethanol elution is collected 50% eluent, drying, dry thing filters with ethanol, 95% ethanol, n-butyl alcohol or dissolve with methanol, filtrate adds or does not add organic solvent, placement is separated out, and filtration, drying obtain ginsenoside Rb
1
6, according to each described a kind of ginsenoside Rb of claim 1-4
1, its analyzing detecting method is:
Adopt high performance liquid chromatography to carry out check and analysis; Chromatographic column: C
18Reversed phase chromatographic column; Chromatographic condition and system suitability experiment: with the octadecylsilane chemically bonded silica is filler; Flow velocity 1.0ml/min; 25 ℃ of column temperatures; Detect wavelength 203nm; Number of theoretical plate is by ginsenoside Rb
1Meter should be not less than 2500; With water: acetonitrile volume ratio=33:67 is a mobile phase;
The preparation of reference substance solution: precision takes by weighing ginsenoside Rb
1Reference substance adds dissolve with methanol and shakes up in volumetric flask, and is diluted to scale;
The preparation of need testing solution: precision takes by weighing or measures sample and adds dissolve with methanol in the volumetric flask and shake up, and is diluted to scale;
Algoscopy: accurate respectively absorption reference substance solution and need testing solution respectively inject chromatograph of liquid, adopt by external standard method with calculated by peak area, promptly.
7, according to each described a kind of ginsenoside Rb of claim 1-4
1Pharmaceutical preparation for the unit dose of active substance preparation.
8, a kind of ginsenoside Rb according to claim 5
1, wherein nonpolar the or low pole macroporous resin described in the preparation method is HPD-100, HPD-100A, HPD-300, HPD-400, HPD-400A, HPD-450, D101,1300-I, 1400 or AB-8.
9, a kind of ginsenoside Rb according to claim 5
1, wherein the organic solvent described in the preparation method is one or both in acetone, the ethyl acetate.
10, a kind of ginsenoside Rb according to claim 7
1Be the pharmaceutical preparation of active substance preparation, wherein unit dose is 1mg-5000mg.
11, a kind of ginsenoside Rb according to claim 7
1Be the pharmaceutical preparation of active substance preparation, wherein unit dose is 5mg-2000mg.
12, according to each described a kind of ginsenoside Rb of claim 1-4
1Treat and/or prevent application in hyperlipidemia, cardiovascular and cerebrovascular disease, tumor, aging, the hypomnesis medicine in preparation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2007101219158A CN101390870A (en) | 2007-09-18 | 2007-09-18 | Ginsenoside Rb1 containing impurity ginsenoside Rc |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2007101219158A CN101390870A (en) | 2007-09-18 | 2007-09-18 | Ginsenoside Rb1 containing impurity ginsenoside Rc |
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| Publication Number | Publication Date |
|---|---|
| CN101390870A true CN101390870A (en) | 2009-03-25 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2007101219158A Pending CN101390870A (en) | 2007-09-18 | 2007-09-18 | Ginsenoside Rb1 containing impurity ginsenoside Rc |
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| CN (1) | CN101390870A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102048778A (en) * | 2009-11-05 | 2011-05-11 | 天津天士力现代中药资源有限公司 | Method for detecting ginseng extract |
| US20150126463A1 (en) * | 2013-11-04 | 2015-05-07 | Hong Kong Baptist University | Use of herbal saponins to regulate gut microflora |
-
2007
- 2007-09-18 CN CNA2007101219158A patent/CN101390870A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102048778A (en) * | 2009-11-05 | 2011-05-11 | 天津天士力现代中药资源有限公司 | Method for detecting ginseng extract |
| CN102048778B (en) * | 2009-11-05 | 2013-10-16 | 天津天士力现代中药资源有限公司 | Method for detecting ginseng extract |
| US20150126463A1 (en) * | 2013-11-04 | 2015-05-07 | Hong Kong Baptist University | Use of herbal saponins to regulate gut microflora |
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Application publication date: 20090325 |